Binding and phosphorylation of tubulin by G protein-coupled receptor kinases

Although the beta-adrenergic receptor kinase (betaARK) mediates agonist-dependent phosphorylation and desensitization of G protein-coupled receptors, recent studies suggest additional cellular functions. During our attempts to identify novel betaARK interacting proteins, we found that the cytoskeletal protein tubulin could specifically bind to a betaARK-coupled affinity column. In vitro analysis demonstrated that betaARK and G protein-coupled receptor kinase-5 (GRK5) were able to stoichiometrically phosphorylate purified tubulin dimers with a preference for beta-tubulin and, under certain conditions, the betaIII-isotype. Examination of the GRK/tubulin binding characteristics revealed that tubulin dimers and assembled microtubules bind GRKs, whereas the catalytic domain of betaARK contains the primary tubulin binding determinants. In vivo interaction of GRK and tubulin was suggested by the following: (i) co-purification of betaARK with tubulin from brain tissue; (ii) co-immunoprecipitation of betaARK and tubulin from COS-1 cells; and (iii) co-localization of betaARK and GRK5 with microtubule structures in COS-1 cells. In addition, GRK-phosphorylated tubulin was found preferentially associated with the microtubule fraction during in vitro assembly assays suggesting potential functional significance. These results suggest a novel link between the cytoskeleton and GRKs that may be important for regulating GRK and/or tubulin function.     

Intrathecal vincristine: an analysis of reasons for recurrent fatal chemotherapeutic error with recommendations for prevention

Activated protein C (APC) is a naturally occurring anticoagulant that interacts with factor V and VIII to inhibit the clotting cascade. The prevalence of APC resistance among Korean patients with deep vein thrombosis is ill defined. The aim of the present study was to investigate the prevalence of APC resistance and factor V Leiden mutation in Korean patients with deep vein thrombosis. The presence of factor V Leiden mutation was determined in 49 patients who visited Asan Medical Center. APC ratio was performed in 33 individuals from the above 49 patients. Three patients were excluded from the analysis because their baseline aPTT was prolonged. Resistance to APC was diagnosed when the APC ratio was below 2.55. APC resistance was documented in 8 individuals, representing 27% (8/30) of the patients on whom APC resistance test was performed. The 2 patients, who showed APC resistance, were positive for lupus anticoagulant. None of the 49 patients demonstrated factor V Leiden mutation. These findings indicate that factor V Leiden mutation is rare and APC resistance is less prevalent in Korean patients with deep vein thrombosis than in Caucasians. APC resistance not caused by factor V Leiden mutation may be a risk factor for deep vein thrombosis in this population.     

INFLUENCE OF AXISYMMETRIC DEFORMATION PROCESSING METHODS ON THE MECHANICAL PROPERTIES OF Cu-Nb COMPOSITES

It has been shown that deformation processed Cu-19% Nb alloys with good strength and electrical conductivity can be developed in sizes that are useful for engineering applications. Mot extrusion of bundled sub-elemental Cu-19% Nb wires followed by cold drawing to make a composite wire of diameter equal to that of the initial sub-elemental wires resulted in a 67% increase in the ultimate tensile stress. However, on subsequent cold drawing of this composite wire the strength increased at a slower rate than that obtained on continuing cold drawing of the sub-elemental wire and the strength differential decreased. In addition, after cold drawing to equivalent diameters the electrical conductivity of the composite wire was less than that of the sub-elemental wire. These results indicate that while high strengths and good electrical conductivities can be produced in larger size deformation processed Cu-Nb composites by a process of bundling, extrusion and cold drawing of sub-elemental wires, there appears to be a limit to the amount of subsequent cold drawing feasible before the benefit in properties ceases.     

Glottal source estimation using a sum-of-exponentials model

An algorithm for estimating the glottal source waveform in voiced speech is described. The glottal source waveform is described using the LF model proposed by Fant et al. (1985). The vocal tract filter is modeled as a pole-zero system. The analysis of vowel sounds from several talkers shows that the analysis procedure leads to an accurate estimate of the glottal source     

Religious ritual and dissociation in India and Australia

Bone metastases, together with generalized bone resorption, represent the main complication in patients with advanced prostate cancer, and palliative treatments are required to delay the progression of the metastases and improve the quality of life of these patients. For this reason, the bisphosphonate clodronate was administered to 18 patients (clodronate group) from a total of 30, all of whom were receiving complete androgenic blockade; the remaining 12 formed the control group. Transiliac bone biopsies were taken at the beginning of the study and 6 months later to determine the effect of the bisphosphonate on the skeleton. The results were assessed by bone histomorphometry and showed, although without statistical significance between the groups, an antiresorptive effect of the clodronate expressed as the eroded surface/bone surface and as the osteoclast number/bone surface. However, the bone volume also decreased after 6 months of treatment. Similarly, osteoid formation decreased as indicated by the osteoid surface and by the osteoid volume, probably due to the effect of the drug on the osteoblasts. The mineralization rate was apparently slightly retarded in the clodronate group, although to a lesser degree than in the control group. The results confirm the antiresorptive effect of clodronate and its detrimental effect on osteoblast activity.     

Robust continuous-time smoothers without two-sided stochastic integrals

We consider the problem of fixed-interval smoothing of a continuous-time partially observed nonlinear stochastic dynamical system. Existing results for such smoothers require the use of two-sided stochastic calculus. The main contribution of the paper is to present a robust formulation of the smoothing equations. Under this robust formulation, the smoothing equations are nonstochastic parabolic partial differential equations (with random coefficients) and, hence, the technical machinery associated with two sided stochastic calculus is not required. Furthermore, the robust smoothed state estimates are locally Lipschitz in the observations, which is useful for numerical simulation. As examples, finite dimensional robust versions of the Benes and hidden Markov model smoothers and smoothers for piecewise linear dynamics are derived; these finite-dimensional smoothers do not involve stochastic integrals.     

Characterization of the proliferation state in canine mammary tumors by the standardized AgNOR method with postfixation and immunohistologic detection of Ki-67 and PCNA

Recently, there have been some reports that changes in serum lipid composition may be related to suicide, major depression and immune-inflammatory responses. Findings from our laboratory suggest that major depression is accompanied by reduced formation of cholesteryl esters and perhaps by impairment of reverse cholesterol transport. The latter is reportedly accompanied by lower serum high-density lipoprotein cholesterol (HDL-C). The aim of this study was to examine whether (i) major depression is accompanied by lower serum HDL-C or by abnormal levels of serum total cholesterol, triglycerides, low-density lipoprotein-C (LDL-C) or vitamin E, (ii) suicidal attempts are related to lower serum HDL-C and (iii) there are significant associations between serum HDL-C and immune/inflammatory markers. A total of 36 subjects with major depression, of whom 28 patients showed treatment resistance, as well as 28 normal control subjects, had blood sampled for the assay of the above lipids, serum zinc (Zn), albumin (Alb) and flow cytometric determination of the T-helper/T-suppressor (CD4+/CD8+) T-cell ratio. In total, 28 depressed subjects had repeated measures of these variables both before and after treatment with antidepressants. Serum HDL-C and total cholesterol, as well as the HDL-C/cholesterol ratio, were significantly lower in subjects with major depression than in normal controls. Serum HDL-C levels were significantly lower in depressed men who had at some time made serious suicidal attempts than in those without such suicidal behaviour. Treatment with antidepressants for 5 weeks did not significantly alter either serum HDL-C or other lipid variables. Serum HDL-C levels were significantly and negatively correlated with the (CD4+/CD8+) T-cell ratio, and positively correlated with serum Alb and Zn. These results suggest that (i) lower serum HDL-C levels are a marker for major depression and suicidal behaviour in depressed men, (ii) lower serum HDL-C levels are probably induced by the immune/inflammatory response in depression and (iii) there is impairment of reverse cholesterol transport from the body tissues to the liver.